For CD1d tetramer immunofluorescence, fresh thymic lobes, whole lymph nodes or 1–2 mm thickness spleen trans-sections were overnight incubated in 50μl phosphate buffered saline (PBS) solution containing 1:10~1:25 dilution of R-Phycoerythrin (PE) labeled PBS-57 loaded CD1d tetramer in 2% fetal calf serum containing PBS at 4°C. Next day, tissues were washed three times with PBS, fixed with 4% paraformaldehyde (PFA) for one hour and snap frozen. Five micrometer tissue sections were blocked with 5% bovine serum albumin and goat sera (Jackson Laboratory) for 1 hour at 25°C and stained with anti-PE antibody (Novus Biologicals, rabbit polyclonal) followed by goat anti-rabbit AF555 (Life Technology). For human CD2 immunofluorescence, tyramide based amplification was done according to manufacturer’s instruction (PerkinElmer) after staining with anti-human CD2 (RPA2) antibody. RORγt (RORg2, Millipore or Q31-378, BD) and PLZF (D9, SantaCruz or R17-809, BD) were stained followed by goat anti-hamster IgG (Jackson Laboratory) and goat anti-mouse IgG1 (Southern Biotech).
R17 809
Manufactured by BD
The R17-809 is a laboratory equipment manufactured by BD. It is designed for general laboratory use, but a detailed description of its core function is not available without the risk of making unsubstantiated claims or interpretations.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti rabbit af555, by Thermo Fisher Scientific (1 mentions)
Goat anti hamster igg, by Jackson ImmunoResearch (1 mentions)
Q31 378, by BD (1 mentions)
Bovine serum albumin (bsa), by Jackson ImmunoResearch (1 mentions)
Goat sera, by Jackson ImmunoResearch (1 mentions)
Anti pe antibody, by Novus Biologicals (1 mentions)
Goat anti mouse igg1, by Southern Biotech (1 mentions)
Ncam16, by BD (1 mentions)
Immu510, by Beckman Coulter (1 mentions)
Fixable live dead stain, by Thermo Fisher Scientific (1 mentions)
2 protocols using r17 809
1
Multiparametric Immunofluorescence Staining
2
Multiparametric Flow Cytometry Analysis
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For phenotypic analysis of lymphocyte subsets by multi-color flow cytometry, PBMCs were stained with fluorochrome-conjugated antibodies for the surface markers CD3 (OKT3, Biolegend), CD4 (SK3, Biolegend), CD8 (SK1, Biolegend), CD14 (M5E2, Biolegend), CD19 (HIB19, Biolegend), CD56 (NCAM16.2, BD Biosciences), CD94 (DX22, Biolegend), CD161 (HP-3G10, Biolegend), CRACC (235614, R&D Systems) NKG2A (Z199, Beckman Coulter), NKp80 (MA152, Beckman Coulter), and TCRγδ (IMMU510, Beckman Coulter) as well as for intracellular EOMES (WD1928, eBioscience), granulysin (GNLY) (DH2, Biolegend), NKG7 (2G9, Beckman Coulter), and PLZF (R17-809, BD Biosciences). Intracellular HOPX was detected with an unconjugated primary antibody (rabbit polyclonal, Proteintech) followed by a fluorochrome-labeled F(ab)2 anti-rabbit IgG secondary antibody (goat polyclonal, ThermoFisher). Dead cells were identified using a fixable live/dead stain (ThermoFisher).
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